A rapid, easy, and sensitive method for detecting His-tag-containing chitinase based on ssDNA aptamers and gold nanoparticles

Z Cao, S Wang, Z Liu, C Xue, X Mao - Food chemistry, 2020 - Elsevier
Z Cao, S Wang, Z Liu, C Xue, X Mao
Food chemistry, 2020Elsevier
Chitooligosaccharides are oligosaccharides with many biological activities that can be used
in food production for sweeteners, preservatives and humectants, among other products.
Chitin, a long-chain polymer of N-acetylglucosamine and a derivative of glucose, can be
hydrolyzed by applying chitinase to break down glycosidic bonds to form
chitooligosaccharides. Chitinases arising from heterologous gene expression are usually
linked to a 6× His-tag to facilitate easy purification. Heterologously expressed chitinase …
Abstract
Chitooligosaccharides are oligosaccharides with many biological activities that can be used in food production for sweeteners, preservatives and humectants, among other products. Chitin, a long-chain polymer of N-acetylglucosamine and a derivative of glucose, can be hydrolyzed by applying chitinase to break down glycosidic bonds to form chitooligosaccharides. Chitinases arising from heterologous gene expression are usually linked to a 6 × His-tag to facilitate easy purification. Heterologously expressed chitinase linked to a 6 × His-tag is a transgenic element, but enzyme activity tests cannot be used to distinguish transgenic elements from natural elements. In this study, we established a rapid and easy method to detect His-tag-containing chitinase using gold nanoparticles (AuNPs) and ssDNA aptamers. Using this method, His-tag-containing chitinase could be detected at concentrations as low as 0.136 nM within 5 min. Color changes of AuNPs showed a positive correlation with His-tag-containing chitinase concentrations.
Elsevier
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